• Title/Summary/Keyword: Division of Fraction

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Effects of Fertilization Time and Culture Medium of Pig Oocytes Matured In Vitro by liquid Boar Sperm Stored at $4^{\circ}C$ (체외성숙된 돼지난포란을 $4^{\circ}C$ 보존 액상정액으로 체외수정시 수정시간과 배양배지의 영향)

  • Park, C. S.;Y. J. Yi;Kim, M. Y.;Y. J. Chang;Lee, S. H.;D. I. Jin
    • Korean Journal of Animal Reproduction
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    • v.27 no.3
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    • pp.215-223
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    • 2003
  • This study was to investigate the effects of fertilization time and culture medium of pig oocytes matured in-vitro by liquid boar sperm. The sperm rich fraction (30∼60 ml) was slowly cooled to room temperature (20∼23$^{\circ}C$) by 2 h after collection. Semen was transferred into 15 ml tubes, centrifuged at room temperature for 10 min 800 ${\times}$ g, and the supernatant solution was poured off. The concentrated sperm was resuspended with 5 ml of the LEN diluent to provide 1.0${\times}$10$^{9}$ sperm/ml at room temperature. The resuspended semen was cooled in a refrigerator to 4$^{\circ}C$. The medium used for oocyte maturation was TCM-199 supplemented with 26.19 mM sodium bicarbonate, 0.9 mM sodium pyruvate, 10 $\mu\textrm{g}$/ml insulin, 2 $\mu\textrm{g}$/ml vitamin B$_{12}$ , 25 mM HEPES, 10 $\mu\textrm{g}$/ml bovine apotransferrin, 150 $\mu$M cysteamine, 10 IU/ml PMSG, 10 IU/ml hCG, 10 ng/ml EGF, 0.4% BSA, 75 $\mu\textrm{g}$/ml sodium penicillin G, 50 $\mu\textrm{g}$/ml streptomycin sulfate and 10% pFF. After about 22 h of culture, oocytes were cultured without cysteamine and hormones for 22 h at 38.5$^{\circ}C$, 5% $CO_2$ in air. Oocytes were inseminated with liquid boar sperm stored at 4$^{\circ}C$ for 2 days after collection. Oocytes were coincubated for 1, 3, 6 and 9 h in 500 ${mu}ell$ mTBM fertilization media with 1.0${\times}$10$^{6}$ sperm/ml concentration, respectively. Thereafter, oocytes were transferred into 500 ${mu}ell$ NCSU-23, HEPES buffered NCSU-23, PZM-3 and PZM-4 culture media, respectively, for further culture of 6, 48 and 144 h. The rates of sperm penetration and male pronuclear formation were higher in the fertilization times for 6 and 9 h than in those for 1 and 3 h. The rates of cleaved oocytes were higher in the fertilization times for 6 and 9 h (85.0 and 84.6%) than in those for 1 and 3 h (61.1 and 76.8%). The percentage of blastocyst formation from the cleaved oocytes was highest in the fertilization time for 6 h (33.6%) than in that for 1, 3 and 9 h (11.4, 23.0 and 29.6%). Mean cell numbers per blastocyst were 32.9, 27.6, 26.3 and 24.4 in the fertilization times for 6, 9, 3 and 1 h, respectively. The rate of blastocyst from the cleaved oocytes and the number of cells per blastocyst were higher in HEPES buffered NCSU-23 culture medium than in NCSU-23, PZM-3 and PZM-4 culture media. In conclusion, we found out that liquid boar sperm stored at 4$^{\circ}C$ could be used for in-vitro fertilization of pig oocytes matured in-vitro. Also, we recommend the coincubation time of 6 h in 500 ${mu}ell$ TBM fertilization medium with 1${\times}$10$^{6}$ sperm/ml concentration and the HEPES buffered NCSU-23 culture medium for in-vitro fertilization of pig oocytes matured in-vitro.

Anti-tumor Effect of Combined Betacarotene with X-irradiation in the Mouse Fibrosarcoma : Cytotoxicity and Tumor Growth Delay (쥐 섬유육종에서 베타카로틴과 방사선조사 병용의 항종양 효과: 세포독성 및 종양성장 지연에 미치는 영향)

  • Kwon Hyoung-Cheol;Yang Moon-Sik
    • Radiation Oncology Journal
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    • v.18 no.2
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    • pp.133-137
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    • 2000
  • Purpose :To investigate whether combined beta-carotene with X-Irradiation has more enhanced radition response than X-irradiation or not, we peformed a experiment about in vitro cytotoxlcity of beta-carotene and/or X-irradiation in the fibrosarcoma cells, tumor growth delay of combined beta-caroten with/or X-irradiation in the mouse fibrosarcoma. Materials and Methods : 2$\%$ emulsion of beta-carotene was serially diluted and used. X-Irradiation was given by 6 MeV linear accelerator. The cytotoxicity of beta-carotene in vitro was evaluated from clonogenic assay. To compare the cytotoxiclty between combined beta-carotene with X-irradiation and X-irradiation group, 2 mg/ml of beta-carotene was contacted to fibrosarcoma (FSall) cells for 1 hour before X-irradiation. For the tumor growth delay, single 20 Gy was given to FSall tumor hearing C3H/N mice whic was classified as beta-crotene with X-irradiation group (n=5) and X-irradiation alone group (n=5). 0.2 ml of 20 mg/kg of beta-carotene were i.p. injected to mice 30 minute before X-irradiation in the beta-crotene with X-irradiation group. The tumor growth delay defined as the time which reach to 1,000 mm$^{3}$ of tumor volume. Results : (1) Cytotoxicity in vitro: 1) survival fraction at beta-carotene concentration of 0.002,0.02,0.2 and 2 mg/ml were 0.69$\pm$0.07, 0.59$\pm$0.08, 0.08$\pm$0.008 and 0.02$\pm$0.006, respectively. 2) each survival fraction at 2, 4, 6 and 8 Gy in the 2 mg/ml of beta-carotene + X-irradiation group were 0.13$\pm$0.05, 0.03$\pm$0.005, 0.01 $\pm$0.002 and 0.009$\pm$0.0008, respectively. But each survival fraction at same irradiation dose in the X-irradiation group were 0.66$\pm$0.05, 0.40$\pm$0.04, 0.11$\pm$0.01 and 0.03$\pm$0.006, respectively(p<0.05). (2) The time which reach to 1,000 mm$^{3}$ of tumor volume of beta-carotene + X-irradiation group and X-irradiation alone group were 18, 19 days, respectively(p>0.05) Conclusion : The contact of beta-caroten to Fsall cells showed mild cytotoxicity which 띤as increased according to concentration. The cytotoxicity of combined beta-carotene with X-irradiation more increased than that of X-irradiation, additionally, And there was significant difference of cytotoxicity between two groups. But there were no significant difference of the growth delay of fibrosarcoma between two groups.

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20 Years Surgical Experiences for Ebstein's Anomaly (엡스타인 기형의 20년 수술 치험)

  • Lee, Sak;Park, Han-Ki;Lee, Chang-Young;Chang, Byung-Chul;Park, Young-Hwan
    • Journal of Chest Surgery
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    • v.40 no.4 s.273
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    • pp.280-287
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    • 2007
  • Background: We retrospectively evaluated the clinical results of surgically managing patients with Ebstein's anomaly. Material and Method: Between Feb. f 984 and June 2006, 50 patients who underwent surgical treatment for Ebstein's anomaly at Yonsei Cardiovascular Center were retrospectively reviewed. The mean age of the patients was 26.9 years and 19 patients were male, Associated anomalies included atrial septal defect (33), patent ductus arteriosus (2), ventricular septal defect (1), and pulmonary stenosis (4), and 90%, (45/50) of the patients had more than a moderate degree of tricuspid regurgitation. Carpentier type A was present in 6 patients, type B in 26, type C in 14 and type D in 4. Ten patients were associated with WPW syndrome. Conservative surgery was possible in 31 patients (tricuspid annuloplasty, plication of the atrialized RV), Fontan's operation was peformed in 4 patients, tricuspid valve replacement was done in 12 and palliative surgery was done in 2 patients. Thirteen patients were associated with hi-directional cavopulmonary shunt (BCPS: one and a half ventricular repairs): 10 patients with WPW syndrome and 4 patients with atrial fibrillation underwent concomitant ablation. Result: The postoperative median NYHA functional class $(3{\rightarrow}1)$ and the mean cardio-thoracic ratio $(0.65{\rightarrow}0.59)$ were decreased significantly (p<0.001, p=0.014). The mean oxygen saturation $(86.6{\rightarrow}94.1%)$, and median TR grade $(4{\rightarrow}1)$ were also significantly improved (p=0.004, p<0.001). For comparison of BCPS and conservative surgery, the preoperative right ventricular pressure (33.0 vs. 41.3 mmHg), the ICU stay (2.80 vs. 1.89 days), the hospital say (10.6 vs. 16.8 days), and the left ventricular ejection fraction (64.3 vs. 72.8%) were statistically different. Postoperative mortality occurred in 3 patients (6%) due to biventricular failure in 2 patients and sepsis in the other patient. The mean follow up duration was 101.5 months, and one patient died of Fontan failure and 6 patients required reoperation (bioprosthetic degenerative change (2) and Fontan conversion (4)). The overall survival rate at 10 years was 90.2%, the freedom from reoperation rate and rate of cardiac related events were 78.9% and 49.2%, respectively. Conclusion: Surgical management of Ebstein's anomaly can be performed safely, and the associated BCPS may be helpful for high-risk patients. Adequate application of surgical management may increase the long-term survival with a reduced rate of reoperation.

Effects of Organic Amendments on Introducing Pioneer Herbaceous Plants in the Abandoned Zinc Mine Soil Revegetation (아연 폐광산에 식생도입을 위한 유기성 토양 개량제의 처리효과)

  • Kim Dae-Yeon;Lee Sang-Hwan;Jung Jin-Ho;Kim Jeong-Gyu
    • Journal of Soil and Groundwater Environment
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    • v.11 no.3
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    • pp.43-51
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    • 2006
  • Generally abandoned mine soils have serious problems for introducing vegetation such as nutrient deficiency, poor physical properties, and phytotoxicity due to high levels of heavy metals. It is required to improve soil amenity for revegetation. One of its strategies is using organic materials such as compost manure and sludge. The pot experiments was conducted to evaluate the effects of pig manure and municipal sewage sludge on revegetation of mining area soil surface with Artemisia princeps and Zoysia japonica. Application rate of pig manure and municipal sewage sludge was $75{\sim}225$ Mg/ha and $150{\sim}450$ Mg/ha, respectively. The results showed that the application of manure and sludge increased organic matter about two-fold and total nitrogen contents about five-fold of mine soil and improved the growth of plants in all treatments compared to the control. The result of plant tissue analysis showed that both plants accumulate Cd, Cu and Zn in root tissue rather than shoot tissues. Increased sludge application reduced Zn accumulation in both plant tissue. Sequential extraction results indicated that addition of soil amendment induced increment of organically bound fractions of Cu and Zn. Organically bound fraction of Zn was significantly increased from 7.84% to 13.58% in Artemisia princeps planted soil and from 7.84% to 14.16% in Zoysia japonica planted soil, thereby bioavailability of heavy metals was reduced. The results suggested that application of organic materials to mine soil can reduce phytotoxicity of heavy metals and be helpful in introducing successful revegetation.

The Influences of Coastal Upwelling on Phytoplankton Community in the Southern Part of East Sea, Korea (동해 남부 연안 해역에서 냉수대 발생이 식물플랑크톤 군집에 미치는 영향)

  • Kim, A-Ram;Youn, Seok-Hyun;Chung, Mi-Hee;Yoon, Sang-Chol;Moon, Chang-Ho
    • The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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    • v.19 no.4
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    • pp.287-301
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    • 2014
  • In order to understand environment condition and phytoplankton community before and after coastal upwelling, the influences of upwelling events on phytoplankton community were studied at 18 stations located the Southern part of East Sea, Korea from May to August 2013. The surface water masses showed low temperature and high salinity due to upwelling events at coastal stations (A1, B1, C1). Correlation between temperature and nutrients (DIP, r=-0.218, p<0.01; DIN, r=-0.306, p<0.01; silicate, r=-0.274, p<0.01) was significantly negative. This result could be explained that nutrients were supplied to surface water by the upwelling of bottom water. Phytoplankton communities were composed of 186 species. Phytoplankton abundance were relatively high in May (C1, $726{\times}10^3cells\;L^{-1}$) and July (A1, $539{\times}10^3cells\;L^{-1}$). Total chlorophyll a and micro-size fraction ($&gt;20{\mu}m$) increased at coastal stations in July and August, while phytoplankton abundance and total chl. a was much low in June. Dominant species in June was Pseudo-nitzschia spp. of which the cell size was $309{\mu}m^3$. Cell size of Pseudo-nitzschia spp. was smaller than dominant species in other period. Therefore, the increase in total chloro-phyll a and the size of phytoplankton was resulted in the sufficient supply of nutrients. In contrast, these tendencies were not observed at outside stations. These results suggested that coastal upwelling was an important influencing factor to determine the species composition and standing stock of phytoplankton community in the coastal waters of the Southern part of East Sea, Korea.

Clay Mineralogical Characteristics and Origin of Sediments Deposited during the Pleistocene in the Ross Sea, Antarctica (남극 로스해 대륙대 플라이스토세 코어 퇴적물의 점토광물학적 특성 및 기원지 연구)

  • Jung, Jaewoo;Park, Youngkyu;Lee, Kee-Hwan;Hong, Jongyong;Lee, Jaeil;Yoo, Kyu-Cheul;Lee, Minkyung;Kim, Jinwook
    • Journal of the Mineralogical Society of Korea
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    • v.32 no.3
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    • pp.163-172
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    • 2019
  • A long core (RS15-LC48) was collected at a site in the continental rise between the Southern Ocean and the Ross Sea (Antarctica) during the 2015 Ross Sea Expedition. The mineralogical characteristics and the origin of clay minerals in marine sediments deposited during the Quaternary in the Ross Sea were determined by analyzing sedimentary facies, variations in grain size, sand fraction, mineralogy, clay mineral composition, illite crystallinity, and illite chemical index. Core sediments consisted mostly of sandy clay, silty clay, or ice rafted debris (IRD) and were divided into four sedimentary facies (units 1-4). The variations in grain size distribution and sand content with depth were very similar to the variations in magnetic susceptibility. Various minerals such as smectite, chlorite, illite, kaolinite, quartz, and plagioclase were detected throughout the core. The average clay mineral composition was dominated by illite (52.7 %) and smectite (27.7 %), with less abundant clay minerals of chlorite (11.0 %) and kaolinite (8.6 %). The IC and illite chemical index showed strong correlation trends with depth. The increase in illite and chlorite content during the glacial period, together with the IC and chemical index values, suggest that sediments were transported from the bedrocks of the Transantarctic Mountains. During the interglacial period, smectite may have been supplied by the surface current from Victoria Land, in the western Ross Sea. High values for IC and the illite chemical index also indicate relatively warm climate conditions during that period.

Temporal and Spatial Characteristics of Sediment Yields from the Chungju Dam Upstream Watershed (충주댐 상류유역의 유사 발생에 대한 시공간적인 특성)

  • Kim, Chul-Gyum;Lee, Jeong-Eun;Kim, Nam-Won
    • Journal of Korea Water Resources Association
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    • v.40 no.11
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    • pp.887-898
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    • 2007
  • A physically based semi-distributed model, SWAT was applied to the Chungju Dam upstream watershed in order to investigate the spatial and temporal characteristics of watershed sediment yields. For this, general features of the SWAT and sediment simulation algorithm within the model were described briefly, and watershed sediment modeling system was constructed after calibration and validation of parameters related to the runoff and sediment. With this modeling system, temporal and spatial variation of soil loss and sediment yields according to watershed scales, land uses, and reaches was analyzed. Sediment yield rates with drainage areas resulted in $0.5{\sim}0.6ton/ha/yr$ excluding some upstream sub-watersheds and showed around 0.51 ton/ha/yr above the areas of $1,000km^2$. Annual average soil loss according to land use represented the higher values in upland areas, but relatively lower in paddy and forest areas which were similar to the previous results from other researchers. Among the upstream reaches, Pyeongchanggang and Jucheongang showed higher sediment yields which was thought to be caused by larger area and higher fraction of upland than other upstream sub-areas. Monthly sediment yields at the main outlet showed same trend with seasonal rainfall distribution, that is, approximately 62% of annual yield was generated during July to August and the amount was about 208 ton/yr. From the results, we could obtain the uniform value of sediment yield rate and could roughly evaluate the effect of soil loss with land uses, and also could analyze the temporal and spatial characteristics of sediment yields from each reach and monthly variation for the Chungju Dam upstream watershed.

Studies on Bovine Serum Protein Fractions (I, II) (혈청단백질분획(血淸蛋白質分劃)에 대한 연구(硏究) (I, II))

  • Rim, Bong Ho
    • Korean Journal of Veterinary Research
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    • v.5 no.1
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    • pp.1-16
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    • 1965
  • I. A Comparison of Sodium Sulfate Precipitation and Zone(Paper, Agar) Electrophoresis; Many kinds of techniques have been used for fractionating serum proteins. In the present study, using bovine serum, the fractions obtained with sodium sulfate were compared with those determined by zone electrophoresis. 1. Fibrinogen was precipitated with 4 to 10 percent of sodium sulfate. 2. ${\gamma}$-globulin required 10 to 16 percent of the salt for precipitation. 3. ${\beta}$-globulin began to precipitate at 12 percent sodium sulfate, and completed precipitation at approximately 26 percent in paper electrophoresis, while at 22 percent in agar electrophoresis. 4. ${\alpha}$-globulin completed precipitation at 13 to 28 percent sodium sulfate in paper electrophoresis and at 22 percent in agar electrophoresis. 5. Albumin began to precipitate at 14 percent of the salt, and was free from the mixture of globulins approximately at 28 percent in paper electrophoresis, while at 22 percent in agar electrophoresis. The results of comparing fractions by the two methods were as follows: 1. Euglobulin (15%) was equal to the sum of the most ${\gamma}$-globulin and a small quantity of the ${\alpha}$-, and ${\beta}$-globulins. 2. Pseudoglobulin I (15-17.5%) corresponded to the most ${\alpha}$-, ${\beta}$-globulins and a small quantity of albumin. 3. Pseudoglobulin II(18-22%) was a mixture of the ${\alpha}$-, ${\beta}$-globulins and albumin fraction. 4. Albumin (above 22%) contained the most albumin fraction separated by zone electrophoresis and a small quantity of the ${\alpha}$-, and ${\beta}$-globulins. As mentioned above the fractions obtained with sodium sulfate were a mixture of the various proportion of the fractions determined by zone electrophoresis. The solubility of serum fractions to sodium sulfate coincided with the mobility of those by zone electrophoresis. (By percent of sodium sulfate we mean gram of sodium sulfate contained in $100m{\ell}$ of solution). II. Immunological Studies on Serum Protein Fractions with Sodium Sulfate; In the previous report the fractions of bovine serum protein with sodium sulfate compared with those obtained by zone electrophoresis, and the findings were that the former contained various proportion components of the latter. In this study the author studied whether or not the fractions with sodium sulfate are simple component antigenically by immunoelectrophoresis and micro double diffusion test (Immuno-precipitation), using rabbit antiserum to bovine serum. In immunoelectrophoresis, normal bovine serum developed with rabbit antibovine serum showed about ten distinct precipitin arcs. The distribution of these arcs was as follows: 1 albumin, 2 ${\alpha}_1$-, 3 ${\alpha}_2$-, 2 ${\beta}_1$-, ${\beta}_2$-, and 1 ${\gamma}$-globulin (Fig. 7, 9). In micro double diffusion test, five to six precipitation bands could be seen between antigens and antibody, the order of the precipitation bands location is albumin, ${\alpha}$-, ${\beta}$-, and ${\gamma}$-globulin from the side of antiserum well (Fig.19). Frequently the ${\alpha}$-, and ${\beta}$-precipitation bands were separated into two or three precipitation bands, which indicated that these globuline are not a pure component antigenically as shown in immuno-electrophoresis. In both Immunological methods, the two ${\alpha}$-, ${\beta}$-precipitin arcs and bands appeared clear and strong, indicating that the two globulins reacted as strong antigens. The precipitate reaction of ${\gamma}$-globulin was shown at 12 to 16 percent sodium sulfate; ${\beta}$-globulin at 12 to 20 percent; ${\alpha}$-globulin at 12 to 22 percent (immuno-electrophoresis), at 12 to 26 percent (Diffusion); and albumin at above 22 percent. Antigenically euglobulin contained ${\gamma}$-, ${\beta}$-, and ${\alpha}$-globulins, Pseudoglobulin I and Pseudoglobulin II were composed of ${\alpha}$-, and ${\beta}$-globulins, and albumin was a mixture of ${\alpha}$-globulin and albumin determined by zone electrophoresis. The results indicated that the fractions of serum protein obtained by either method were constituents of various proteins antigenically except ${\gamma}$-globulin and albumin by Zone electrophoresis.

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Fractionation and Pollution Index of Heavy Metals in the Sangdong Tungsten Mine Tailings (광미에 존재하는 중금속의 분획화와 오염도 평가)

  • Yang, Jae-E.;Kim, Hee-Joung;Jun, Sang-Ho
    • Korean Journal of Soil Science and Fertilizer
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    • v.34 no.1
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    • pp.33-41
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    • 2001
  • Enormous volumes of mining wastes from the abandoned and closed mines are disposed without a proper treatment in the upper Okdong River basin at Southeastern part of Kangwon Province. Erosion of these wastes contaminates soil, surface water, and sediments with heavy metals. Objectives of this research were to fractionate heavy metals in the mine tailing stored in the Sangdong Tungsten tailing dams and to assess the potential pollution index of each metal fraction. Tailing samples were collected from tailing dams at different depth and analyzed for physical and chemical properties. pH of tailings ranged from 7.3 to 7.9. Contents of total N and organic matter were in the ranges of 3.2~5.5%, and 1.3~9.1%, respectively. Heavy metals in the tailings were higher in the newly constructed tailing dam than those in the old dam. Total concentrations of metals in the tailings were in the orders of Zn > Cu > Pb > Ni > Cd, exceeded the corrective action level of the Soil Environment Conservation Law and higher than the natural abundance levels reported from uncontaminated soils. Relative distribution of heavy metal fractions was residual > organic > reducible > carbonate > adsorbed, reversing the degree of metal bioavailability. Mobile fractions of metals were relatively small compared to the total concentrations. Distribution of metals in the tailing dam profiles was metal specific. Concentrations of Cu at the surface of tailing dams were higher than those at the bottom. Pollution index (PI) values of each fraction of metals were ranged from 4.27 to 8.51 based on total concentrations. PI values of mobile fractions were lower than those of immobile fractions. Results on metal fractions and PI values of the tailing samples indicate that tailing samples were contaminated with heavy metals and had potential to cause a detrimental effects on soil and water environment in the lower part of the stream. A prompt countermeasure to prevent surface of tailings in the dams from water and wind erosions is urgently needed.

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Effect of Solvent Fractions from Methanol Extract of Doenjang on Inhibition of Growth and DNA Synthesis of Human Cancer Cells. (인체 암세포 성장 및 DNA 합성 억제에 미치는 된장 분획물의 영향)

  • LIM Sun-Young;Rhee Sook-Hee;Park Kun-Young
    • Journal of Life Science
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    • v.15 no.5 s.72
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    • pp.685-691
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    • 2005
  • Growth and DNA synthesis inhibitory effects of doenjang methanol extract and its solvent fractions on AGS human gastric adenocarcinoma cells, Hep 3B human hepatocellular carcinoma cells, HT-29 human colon cancer cells and MG-63 human osteosarcoma cells were studied. The treatment of doenjang methanol extract ($ 200{\mu}g/ml $) with the AGS, Hep 3B, HT-29 and MG-63 cancer cells after 6 days of incubation inhibited the growth of cancer cells by $32\%$, $51\%$, $84\%$ and $33\%$, respectively. To separate active compounds of doenjang, doenjang methanol extract was fractionated with dichloromethane, ethylacetate, and buthanol. Among the solvent fractions, the dichloromethane and ethylacetate fractions showed the highest growth inhibitory effects on various cancer cells. For example, the dichloromethane and ethylacetate fractions ($200a{\mu}g/ml$) sig-nificantly inhibited the growth of various cancer cells by $89\∼96\%$ and$62\∼86\%$, respectively. DNA synthesis of AGS and Hep 3B cancer cells was significantly inhibited by adding dichloromethane fraction ($200{\mu}g/ml$) up to $94\%$ and $80\%$, respectively. Similarly, the ethylacetate fraction ($ 200\mug/ml $) showed a $ 95\% $ inhibition rate of DNA synthesis in AGS cells. These results suggest that the dichloromethane and ethylacetate fractions have specific active compounds, which will explain this anticancer effect of doenjang.